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121.
Koichi Rikimaru Hitomi Toda Noriko Tachikawa Nobuyuki Kamata Shoji Enomoto 《In vitro cellular & developmental biology. Plant》1990,26(9):849-856
Summary A novel protein-free synthetic medium has been developed for the culture of human squamous cell carcinoma cells. This medium,
designated PF86-1, supports the serial subcultivation of six out of nine human squamous cell carcinoma cell lines in a protein-free,
chemically defined condition without the adapting culture from serum-containing conditions. These cell lines growing in PF86-1
exhibited nearly equal potency to grow in massive culture without noticeable changes in morphology but presented a significantly
decreased level of colony forming efficiency when compared with the cells cultured in serum-containing media, suggesting the
implication of some autocrine mechanism. Interestingly, this medium supported the growth of normal human squamous cells of
oral mucosa and skin for more than 2 mo. in the primary explant culture in spite of high levels of calcium ion concentration,
where the overgrowth of fibroblasts as contaminant was not observed. These results suggest that PF86-1 supports the growth
of cells derived from epidermal tissues selectively and provides the same defined condition for growth of malignant and nonmalignant
human squamous cells. It seems, therefore, that PF86-1 allows investigations on the products of squamous cell carcinoma cells
or on the differences of growth mechanisms between normal and neoplastic human squamous cells. 相似文献
122.
Anna L. Trifillis Myong Won Kahng 《In vitro cellular & developmental biology. Plant》1990,26(5):441-446
Summary We have developed an in vitro model of human papillary collecting duct cells isolated from cadaver kidneys using methods similar
to those we previously reported for the isolation of human proximal tubule cells. To date we have isolated papillary collecting
duct cells from 100 normal human kidneys. Papillae were dissected and digested in Cellgro containing 400 U/ml collagenase.
Cells were plated on fibronectin-coated culture flasks at a density of 104 live cells/ml in Cellgro supplemented with insulin and 10% fetal bovine serum. Confluent monolayers, which were able to withstand
600 mOsm for 8 h, were obtained within 10 to 15 d. Cells of primary isolates and first passages exhibited epithelial cell ultrastructure
including cell junctions, microvilli, and cilia. A dark-brown reaction product was observed in these cells when stained by
the immunoperoxidase method with peroxidase-labeled peanut lectin (Arachis hypogaea), which binds specifically to human distal tubule and collecting duct cells. These cells were negative for Factor-VIII (a
marker for endothelial cells) and γ-glutamyltransferase (a marker for proximal tubule cells). High activities of the glycolytic
enzyme pyruvate kinase and arginine vasopressin-stimulated cAMP production in these cells are consistent with a distal nephron
origin. The results indicate that human collecting duct cells can be isolated and cultured to provide an in vitro system to
probe pathogenetic mechanisms of potential nephrotoxins.
Part of this work was presented at a Symposium of the Center for Alternatives to Animal Testing, April 4–5, 1989, Johns Hopkins
Medical Institutions, Baltimore, MD 21205.
This work was supported in part by grants R01-AI24179, PO1-A804393 for the Public Health Service, U.S. Department of Health
and Human Services, and by a grant from the National Kidney Foundation, Baltimore, MD affiliate. 相似文献
123.
How to Characterize a Biological Antioxidant 总被引:15,自引:0,他引:15
Barry Halliwell 《Free radical research》1990,9(1):1-32
An antioxidant is a substance that, when present at low concentrations compared to those of an oxidizable substrate, significantly delays or prevents oxidation of that substrate. Many substances have been suggested to act as antioxidants in vivo, but few have been proved to do so. The present review addresses the criteria necessary to evaluate a proposed antioxidant activity. Simple methods for assessing the possibility of physiologically-feasible scavenging of important biological oxidants (superoxide, hydrogen peroxide, hydroxyl radical, hypochlorous acid, haem-associated ferryl species, radicals derived from activated phagocytes, and peroxyl radicals, both lipid-soluble and water-soluble) are presented, and the appropriate control experiments are described. Methods that may be used to gain evidence that a compound actually does function as an antioxidant in vivo are discussed. A review of the pro-oxidant and anti-oxidant properties of ascorbic acid that have been reported in the literature leads to the conclusion that this compound acts as an antioxidant in vivo under most circumstances. 相似文献
124.
T. C. Hsu Edward J. Shillitoe Lorraine M. Cherry Qi Lin Stimson P. Schantz Cynthia Furlong 《In vitro cellular & developmental biology. Plant》1990,26(1):80-84
Summary Forty lymphoblastoid (lymphoid) lines were established from 42 volunteer blood donors, including healthy individuals and patients
with head and neck carcinomas. Each peripheral blood sample was split into two portions, one for the establishment of a lymphoid
line and the other for short-term culture, which was used to estimate bleomycin sensitivity by cytogenetic procedures. Twenty
lymphoid lines were selected at random to compare bleomycin sensitivity with data obtained from short-term lymphocyte cultures.
In each set, bleomycin sensitivity of lymphoid cells was similar to that of the lymphocytes. The lymphoid lines, which can
be propagated for an unlimited supply of relatively homogeneous cellular material, will be useful for a variety of future
investigations.
This investigation was supported by grants from the John S. Dunn Foundation, Houston, TX, the Esther Knispel Fund administered
by The University of Texas M. D. Anderson Cancer Center, Houston, TX, and Department of Health and Human Services PHS grant
DE 07007. 相似文献
125.
M. Couturier F. Lemonnier M. Conti M. Feneant-Thibault A. Lemonnier 《In vitro cellular & developmental biology. Plant》1990,26(1):29-32
Summary 2-Deoxy-d-glucose (2-DOG) uptake was tested in human fibroblast cultures in the presence and absence of vitamin E. Addition of 10 μg/ml
vitamin E to the culture medium significantly reduced this uptake for 2-DOG concentrations of 0.005, to 10 mmol/liter (P≤0.01). The decrease of 2-DOG uptake was inversely proportional to the rise in 2-DOG concentration (P≤0.01). The presence of vitamin E reduced by 71% the average cellular level of lipid peroxides (expressed as thiobarbituric
acid reactive substances) and caused a small but significant decrease in the cholesterol concentration (P≤0.01). These last results might explain the decrease in 2-DOG uptake observed in the presence of vitamin E. 相似文献
126.
The Wolf-Hirschhorn syndrome (WHS) is caused by a partial deletion in the short arm of chromosome 4 band 16.3 (4p16.3). A unique-sequence human DNA probe (39 kb) localized within this region has been used to search for sequence homology in the apes' equivalent chromosome 3 by FISH-technique. The WHS loci are conserved in higher primates at the expected position. Nevertheless, a control probe, which detects alphoid sequences of the pericentromeric region of humans, is diverged in chimpanzee, gorilla, and orangutan. The conservation of WHS loci and divergence of DNA alphoid sequences have further added to the controversy concerning human descent. 相似文献
127.
Toshio Ariga Shama Bhat Takashi Kanda Masanaga Yamawaki Tadashi Tai Yasunori Kushi Takeshi Kasama Shizuo Handa Robert K. Yu 《Glycoconjugate journal》1996,13(2):135-145
We analysed the glycolipid composition of glioma cells (N-370 FG cells), which are derived from a culture of transformed human fetal glial cells. The neutral and acidic glycolipid fractions were isolated by column chromatography on DEAE-Sephadex and analysed by high-performance thin-layer chromatography (HPTLC). The neutral glycolipid fraction contained 1.6 µg of lipid-bound glucose/galactose per mg protein and consisted of GlcCer (11.4% of total neutral glycolipids), GalCer (21.5%), LacCer (21.4%), Gb4 (21.1%), and three unknown neutral glycolipids (23%). These unknown glycolipids were characterized as Lewisx (fucosylneolactonorpentaosyl ceramide; Lex), difucosylneolactonorhexaosyl ceramide (dimeric Lex), and neolactonorhexaosyl ceramide (nLc6) by an HPTLC-overlay method for glycolipids using specific mouse anti-glycolipid antibodies against glycolipid and/or liquid-secondary ion (LSI) mass spectrometry. The ganglioside fraction contained 0.6 µg of lipid-bound sialic acid per mg protein with GD1a as the predominant ganglioside species (83% of the total gangliosides) and GM3, GM2, and GM1 as minor components. Trace amounts of sialyl-Lex and the complex type of sialyl-Lex derivatives were also present. Immunocytochemical studies revealed that GD1a and GalCer were primarily localized on the surface of cell bodies. Interestingly, Lex glycolipids and sialyl-Lex were localized not only on the cell bodies but also on short cell processes. Especially, sialyl-Lex glycolipid was located on the tip of fine cellular processes. The unique localization of the Lex glycolipids suggests that they may be involved in cellular differentiation and initiation of cellular growth in this cell line. 相似文献
128.
Kristina Mrkoci Sørge Kelm Paul R. Crocker Roland Schauer Eric G. Berger 《Glycoconjugate journal》1996,13(4):567-573
Previously, 1,3-galactosyltransferase-deficient (Tn+) and normal (TF+) T-lymphocyte clones have been established from a patient suffering from Tn-syndrome [Thurnheret al. (1992)Eur J Immunol
22: 1835–42], Tn+ T lymphocytes express only Tn antigen (GalNAc1-O-R) while other O-glycan structures such as sialosyl-Tn (Neu5Ac2,6GalNAc1-O-R) or TF (Gal1-3GalNAc1-O-R) antigens are absent from these cells as shown by flow cytometry using specific mABs for TF and sialosyl-Tn antigen, respectively. Normal T lymphocytes express the TF antigen and derivatives thereof. The surface glycans of Tn+ and TF+ cells were then analysed by flow cytometry using the following sialic acid-binding lectins:Amaranthus caudatus (ACA),Maackia amurensis (MAA),Limax flavus (LFA),Sambucus nigra (SNA) andTriticum vulgare (WGA). Equal and weak binding of MAA and SNA to both TF+ and Tn+ cells was found. WGA, LFA and ACA bound more strongly to TF+ cells than to Tn+ cells. Binding of ACA to TF+ cells was enhanced after sialidase treatment. To investigate the possible biological consequences of hyposialylation, binding of three sialic acid-dependent adhesion molecules to Tn+ and TF+ cells was estimated using radiolabelled Fc-chimeras of sialoadhesin (Sn), myelin-associated glycoprotein (MAG) and CD22. Equal and strong binding of human CD22 to both TF+ and Tn+ cells was found. Whereas binding of Sn and MAG to TF+ cells was strong (100%), binding to Tn+ cells amounted only to 33% (Sn) and 19% (MAG). These results indicate that thein vivo interactions of T lymphocytes in the Tn syndrome with CD22 are not likely to be affected, whereas adhesion mediated by Sn or MAG could be strongly reduced. 相似文献
129.
Immaculada Martin Josep A. Villena Marta Giralt Roser Iglesias Teresa Mampel Octavi Vińas Francesc Villarroya 《Molecular and cellular biochemistry》1996,154(2):107-111
The action of thyroid hormones on the expression of the mitochondrial ATP synthase -subunit gene (ATPsyn) is controversial. We detected a binding site for the thyroid hormone receptor between-366 and-380 in the human ATPsyn gene by DNase I footprint analysis and band-shift assays. However, expression vectors in which the chloramphenicol acetyl transferase (CAT) reporter gene is driven by the 5 upstream region of ATPsyn gene were unresponsive to T3 when transiently transfected to HepG2 or GH4C1 cells. CAT constructs driven by the rat phosphoenolpyruvate carboxykinase (PEPCK) or the growth hormone (GH) promoters were stimulated several fold by T3 in parallel experiments. It is proposed that the biological effects of thyroid hormones on the ATPsyn expression occur through indirect mechanisms. 相似文献
130.
Rajinder P. Bhullar 《Molecular and cellular biochemistry》1996,156(1):59-67
A major 27 kDa particulate and a minor 24 kDa cytosolic GTP-binding protein was detected in HEL cells upon incubation with [-32P]GTP of nitrocellulose blots containing polypeptides separated using SDS-PAGE. Addition of lovastatin (30 M) to HEL cells in culture inhibited protein synthesis by 35%. However, this treatment resulted in a 5-fold increase, as quantitated by [-32P]GTP binding, in the amount of cytosolic 24 kDa GTP-binding protein. Addition of cycloheximide plus lovastatin to cells in culture abolished the observed increase in 24 kDa GTP-binding protein. Incubation of cells with lovastatin plus [R,S]-[5-3H]mevalonolactone resulted in the incorporation of radioactivity into several polypeptides in both the cytosolic and particulate fractions including a polypeptide of molecular mass of 24 kDa in the cytosol. The mobility of this 24 kDa isoprenylated protein on SDS-PAGE was identical to that of the GTP-binding protein increased in response to lovastatin. However, the 24 kDa protein remained in the cytosol after undergoing isoprenylation. The 24 kDa protein was distinct from the HEL cell, G25K/CDC42Hs GTP-binding protein and the GTP-binding protein that was a substrate for botulinum toxin C3 catalyzed ADP-ribosylation. Results demonstrate that lovastatin specifically increases the expression of a 24 kDa GTP-binding protein in HEL cells and that, isoprenylation of low molecular mass GTP-binding protein(s) may have function(s) in addition to its role in the targetting of these proteins to cell membrane. 相似文献